Bifunctional Molecules that Induce Both Transcytosis and Degradation of Extracellular Proteins
Date of Award
Spring 2024
Document Type
Dissertation
Degree Name
Doctor of Philosophy (PhD)
Department
Chemistry
First Advisor
Spiegel, David
Abstract
The accumulation of pathogenic proteins is a hallmark of many neurodegenerative diseases, and removal of these species is a promising avenue for the development of novel therapies. Targeted protein degradation technologies are emerging as efficacious therapeutic strategies in a wide range of diseases. However, there are no existing bifunctional molecules capable of crossing the blood-brain barrier and triggering degradation of extracellular proteins in the central nervous system. We present transcytosis-inducing molecular degraders of extracellular proteins (TransMoDEs) capable of target protein removal by both transcytosis and degradation in brain cells. Angiopep-2, a brain-targeting peptide derived from aprotinin, has previously been employed as a covalent tag to facilitate receptor-mediated transcytosis of therapeutics across the blood-brain barrier (BBB). TransMoDEs consisting of Angiopep-2 modified with dinitrophenyl, biotin or a chloroalkane ligand triggered endocytosis of anti-DNP antibody, streptavidin and HaloTag protein, respectively. Interestingly, uptake occurred independently of low-density lipoprotein receptor-related protein 1, the reported receptor for Angiopep-2. TransMoDE-mediated endocytosis of streptavidin in a bEnd.3 BBB model resulted in two mechanisms of protein removal: both bi-directional transcytosis and lysosomal degradation. Next generation compounds involving four covalently-bound Angiopep-2 peptides showed improved uptake as compared to the monomeric Angiopep-2 in brain endothelial cells and hepatocytes. These tetrameric compounds significantly depleted anti-DNP antibody in vivo as compared to vehicle and acetylated controls.
Recommended Citation
Howell, Rebecca Anne, "Bifunctional Molecules that Induce Both Transcytosis and Degradation of Extracellular Proteins" (2024). Yale Graduate School of Arts and Sciences Dissertations. 1326.
https://elischolar.library.yale.edu/gsas_dissertations/1326
